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Separation of abietane-type diterpenoids from Clerodendrum kaichianum Hsu by high-speed counter-current chromatography using stepwise elution
Authors:Mingfeng Xu  Qin Zhu  Huizhong Wang  Qizhen Du
Institution:1. Key Lab of Medical Plant Genetic Improvement and Quality Control of Zhejiang Province, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, China;2. Institute of Food Chemistry, Zhejiang Gongshang University, Hangzhou, China
Abstract:High-speed counter-current chromatography (HSCCC) was successfully used for the separation of abietane-type diterpenoids from the medicinal plant C. kaichianum, which were not separated in our previous study using preparative HPLC. The HSCCC separation employed the lower phases of n-hexane–ethyl acetate–methanol–water (HEMW) 4:5:4:5 and HEMW 4:5:5:4 as the mobile phase for stepwise elution while the upper phase of HEMW 4:5:4:5 was used as the stationary phase. HSCCC separation yielded 90.5 mg of compound 1(kaichianone A), 137.7 mg of compound 2 (kaichianone B), 125.0 mg of compound 3 (teuvincenone E), and 227.6 mg of compound 4 (taxusabietane A) with purities of 95.3%, 97.2%, 97.8%, and 98.6%, respectively, as determined by HPLC. Compounds 12 are two new abietane-type diterpenoids while Compounds 34 are known abietane-type diterpenoids, analyzed by ESIMS and NMR data. The results demonstrated that HSCCC can be an excellent alternative for other separation methods. The two new compounds showed significant cytotoxicity against ileocecal carcinoma HCT-8 and breast adenocarcinoma MCF-7 cells.
Keywords:Abietane-type diterpenoid  Clerodendrum kaichianum Hsu  cytotoxicity  high-speed counter-current chromatography  NMR  stepwise elution
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